LOCUS       sequence-450386-        5863 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein purification construct. Rab11 crystallography truncation 
            with a C-terminal cys for maleimide labeling..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5863)
  TITLE     Harper lab Rab11 and inflammasome reporter plasmids
REFERENCE   2  (bases 1 to 5863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Rab11A[8-175]-His-C
FEATURES             Location/Qualifiers
     source          1..5863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(15..62)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     58..76
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(118..135)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             656..678
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    693..717
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(717..736)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(718..736)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(824..843)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1049..1126
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1127..2209
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1146..1165)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3018..3209
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQTDICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3203..3225)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3311..3450
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3366..3385
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3465..3482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3636..4224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3716..3735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4395..5186,5187..5255))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5018..5037
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5256..5360)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5386..5841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5523..5544)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5735..5754
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
       61 agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttaacagtg
      121 atgatgatgg tgatgttcca gaatgcggta aatctctgtt aaaattgtct gaaaagcagc
      181 ttctacattt gtagagtcta gggccgaagt ttcaatgaat gacaaaccat tcttttctgc
      241 aaaagctctt gcttcatctg taggaactgc cctgagatga cgtagatcac tcttattgcc
      301 cacaagcatg ataacaatgt tactatcagc atgatctctc agttctttca gccatcgctc
      361 tacattttca tatgtgagat gtttagcaat gtcataaacc aataaggcac ctacagctcc
      421 acgataatat gctgatgtta tagctcgata tcgctcttgc cctgctgtgt cccatatctg
      481 tgcctttatt gtttttccat caacctggat gcttcttgtt gcaaactcta ctccaatggt
      541 gctcttgctt tccagattaa actcatttcg agtaaatcga gacaggagat tactctttcc
      601 aacaccagaa tctccaataa ggacaacttt aaagaggtag tcgtacatat gtatatctcc
      661 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
      721 atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta cgccggacgc
      781 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
      841 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
      901 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
      961 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     1021 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
     1081 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
     1141 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
     1201 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
     1261 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
     1321 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
     1381 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
     1441 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
     1501 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
     1561 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
     1621 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
     1681 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
     1741 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
     1801 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
     1861 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
     1921 tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
     1981 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
     2041 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
     2101 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     2161 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     2221 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
     2281 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     2341 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
     2401 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
     2461 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
     2521 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
     2581 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
     2641 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
     2701 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
     2761 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
     2821 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
     2881 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
     2941 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
     3001 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
     3061 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacaga
     3121 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
     3181 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     3241 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     3301 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
     3361 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tgcggtgtga aataccgcac
     3421 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     3481 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3541 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3601 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3661 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3721 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3781 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3841 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3901 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3961 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4021 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4081 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4141 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4201 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4261 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4321 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4381 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4441 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4501 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4561 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4621 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4681 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     4741 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4801 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4861 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4921 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4981 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5041 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5101 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5161 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5221 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5281 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5341 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5401 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5461 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5521 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5581 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5641 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5701 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5761 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5821 atgcgccgct acagggcgcg tcccattcgc caatccggat ata
//
